Source:http://linkedlifedata.com/resource/pubmed/id/15219696
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1-2
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pubmed:dateCreated |
2004-6-28
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pubmed:abstractText |
By combining electron tomography with energy-filtered electron microscopy, we have shown the feasibility of determining the three-dimensional distributions of phosphorus in biological specimens. Thin sections of the nematode, Caenorhabditis elegans were prepared by high-pressure freezing, freeze-substitution and plastic embedding. Images were recorded at energy losses above and below the phosphorus L2,3 edge using a post-column imaging filter operating at a beam energy of 120 keV. The unstained specimens exhibited minimal contrast in bright-field images. After it was determined that the specimen was sufficiently thin to allow two-window ratio imaging of phosphorus, pairs of pre-edge and post-edge images were acquired in series over a tilt range of +/-55 degrees at 5 degrees increments for two orthogonal tilt axes. The projected phosphorus distributions were aligned using the pre-edge images that contained inelastic contrast from colloidal gold particles deposited on the specimen surface. A reconstruction and surface rendering of the phosphorus distribution clearly revealed features 15-20 nm in diameter, which were identified as ribosomes distributed along the stacked membranes of endoplasmic reticulum and in the cytoplasm. The sensitivity of the technique was estimated at < 35 phosphorus atoms per voxel based on the known total ribosomal phosphorus content of approximately 7000 atoms. Although a high electron dose of approximately 10(7)e/nm2 was required to record two-axis tilt series, specimens were sufficiently stable to allow image alignment and tomographic reconstruction.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0304-3991
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pubmed:author | |
pubmed:copyrightInfo |
Copyright 2004 Elsevier B.V.
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pubmed:issnType |
Print
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pubmed:volume |
100
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
115-25
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:15219696-Animals,
pubmed-meshheading:15219696-Caenorhabditis elegans,
pubmed-meshheading:15219696-Freezing,
pubmed-meshheading:15219696-Image Processing, Computer-Assisted,
pubmed-meshheading:15219696-Imaging, Three-Dimensional,
pubmed-meshheading:15219696-Microscopy, Electron,
pubmed-meshheading:15219696-Phosphorus,
pubmed-meshheading:15219696-Sensitivity and Specificity,
pubmed-meshheading:15219696-Specimen Handling,
pubmed-meshheading:15219696-Tomography, X-Ray Computed
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pubmed:year |
2004
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pubmed:articleTitle |
Three-dimensional distributions of elements in biological samples by energy-filtered electron tomography.
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pubmed:affiliation |
Division of Bioengineering and Physical Science, ORS, National Institutes of Health, Building 13, 3N17, Bethesda, MD 20892, USA. leapman@helix.nih.gov
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pubmed:publicationType |
Journal Article,
Comparative Study
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